Concomitant completely ossified trigeminal pore and Dorello's canal.

Commonly, the trigeminal and abducens nerve course to the middle cranial fossa, beneath the tentorial border (posterior petroclinoid dural ligament) and, respectively, beneath Grüber's petrosphenoidal ligament, in Dorello's canal. It is hereby reported a rare unilateral association of anatomic variants which was found when the brain computed tomography angiography of a 56-year-old male patient was observed. On the left side, the tentorial border was ossified above the petrous apex, resulting in a tentorial bar 1.96 cm long that transformed the trigeminal pore into a completely ossified one. On that side was also found an ossified petrosphenoidal ligament determining a completely ossified Dorello's canal. On the opposite side a 2.9 mm long clinoid bar extended from the posterior clinoid process to the anterior one. Although these bars are not common in humans they should be documented in computed tomography in cases with associated trigeminal neuralgia and abducens nerve palsy.

Evidence of lymphatics in the rat eye retina.

BACKGROUND: The lymphatic structure of the eye is still under debate. It is mainly assumed that the retina is primarily drained by prelymphatics and not by lymphatics per se. We aimed to identify lymphatics in the rat retina. METHODS: Eyes from ten Wistar rats were paraffin-embedded and the lymphatic marker podoplanin (D2-40) was investigated. RESULTS: We identified in the rat retina a blunt-end network of lymphatic endothelial vessels. It consisted of circumferential vessels within the outer and, respectively, inner plexiform layers, connected by radial dichotomous vessels. Moreover, D2-40 expression was found within the choroid, ciliary body, and extraocular muscles. CONCLUSIONS: This in situ evidence is strongly supported by the recent in vitro demonstration of the expression of lymphatic markers in retinal endothelial cells. Further studies of comparative histology should use specific lymphatic markers to test whether other species besides rats have proper retinal lymphatics.

Transantral intraseptal sinuous canal.

The sinuous canal is an anatomically well-defined intramural canal of the maxillary sinus (MS) folded within the antral walls. Commonly, its first, infraorbital part, courses within the antral roof, while its second, transverse facial part courses below the infraorbital foramen within the anterior antral wall. While retrospective files of patients that were scanned in cone-beam computed tomography (CBCT) for different dental medical purposes were observed randomly, a peculiar variant of the sinuous canal was noticed and further documented. The respective canal origin was far posterior in the infraorbital groove and the canal coursed through the MS embedded within an incomplete oblique septum dividing the antrum into anterosuperior and posteroinferior chambers. Then the sinuous canal continued with the transverse facial segment. As the sinuous canal contains the superior anterior alveolar nerve and artery, major suppliers of the frontal teeth, it is recommended to document in CBCT a possible transantral, and not intramural, course of it, especially when surgical or endoscopic corridors through the MS are planned.

Bilateral giant and unilateral duplicated sphenoidal tubercle.

The sphenoidal tubercle (SphT), also known as pyramidal tubercle or infratemporal spine projects from the anterior end of the infratemporal crest of the greater sphenoidal wing. As it masquerades the lateral entrance in the pterygopalatine fossa it could obstruct surgical corridors or the access for anaesthetic punctures. The SphT is, however, an overlooked structure in the anatomical literature. During a routine cone beam computed tomography study in an adult male patient we found bilateral giant SphTs transforming the infratemporal surfaces of the greater wing into veritable pterygoid foveae. Moreover, on one side the SphT appeared bifid, with a main giant partition, of 9.17 mm vertical length, and a secondary laminar one. The opposite SphT had 14.80 mm. In our knowledge, such giant and bifid SphTs were not reported previously and are major obstacles if surgical access towards the pterygopalatine fossa and the skull base is intended.

The temporomandibular joint: pneumatic temporal cells open into the articular and extradural spaces.

The pneumatisation of the articular tubercle (PAT) of the temporal squama isa rare condition that modifies the barrier between the temporomandibular joint(TMJ) space and the middle cranial fossa. During a routine examination of thecone-beam computed tomography (CBCT) files of patients who were scannedfor dental medical purposes, we identified a case with multiple rare anatomicvariations. First, the petrous apex was bilaterally pneumatised. Moreover, bilateraland multilocular PAT were observed, while on one side it was further found thatthe pneumatic cells were equally dehiscent towards the extradural space and thesuperior joint space. To the best of our knowledge, such dehiscence has not previouslybeen reported. The two temporomastoid pneumatisations were extendedwith occipital pneumatisations of the lateral masses and occipital condyles, thelatter being an extremely rare evidence. The internal dehiscence of the mandibularcanal in the right ramus of the mandible was also noted. Additionally, doublemental foramen and impacted third molars were found on the left side. Suchmultilocular PAT represents a low-resistance pathway for the bidirectional spreadof fluids through the roof of the TMJ. Further, it could add to a morphologicalpicture of hyperpneumatisation of the posterior cranial fossa floor, which couldsignify the involvement of the last four cranial nerves in the clinical picture of TMJpain.

Telocyte-like cells containing Weibel-Palade bodies in rat lamina fusca.

Telocytes (TCs) are cells with long, thin and moniliform processes called telopodes. These cells have been found in numerous tissues, including the eye choroid and sclera. Lamina fusca (LF), an anatomical structure located at the sclera-choroid junction, has outer fibroblastic lamellae containing cells with long telopodes. The purpose of this study was to evaluate, via transmission electron microscopy, the LF for the presence of endothelial-specific ultrastructural features, such as Weibel-Palade bodies (WPBs), in the residing TCs. We found that the outer fibroblastic layer of LF lacked pigmented cells but contained numerous cells with telopodes. These cells had incomplete or absent basal laminae, were united by focal adhesions and close contacts, and displayed scarce caveolae and shedding vesicles. Within the stromal cells of LF, numerous WPBs in various stages of maturation and vesicular structures, as secretory pods that ensure the exocytosis of WPBs content, were observed. The WPBs content of the cells with telopodes in the LF could indicate either their involvement in vasculogenesis and/or lymphangiogenesis or that they are the P-selectin- and CD63-containing pools that play roles in scleral or choroidal inflammation.

The telopode- and filopode-projecting heterogeneous stromal cells of the human sclera niche.

Telocytes (TCs) are stromal cells defined by the presence of long and slender prolongations (telopodes). They are a biologically and functionally heterogeneous population that has not been previously investigated in the sclera. The purpose of this study is to investigate the presence and characteristics of scleral telocytes through a combined immunohistochemical and transmission electron microscopy (TEM) study using samples from ten adult patients. Stromal cells with a TC-like morphology expressed CD34, CD45, CD105, vimentin and occasionally CD68 but were negative for collagen III, CD31, CD133, and CD146. Conjunctival epithelial cells expressed CD45, CD105, CD146, and vimentin. These phenotypes support a scleral niche with immune TCs and haematopoietic stem cells (HSCs). In TEM, we often found spindle-shaped stromal cells projecting telopodes or filopodes, with extremely long nuclei extended even within those prolongations. We separated these cells into a light subtype, which contained a complete set of organelles, and a dark subtype, consisting of undifferentiated stem/progenitor cells. The light cells contained dense vesicles, Weibel-Palade bodies, and rounded α-granule-like structures. These storage areas for the von Willebrand factor (vWF) are known to express selectins that are critically involved in HSC homing and could also indicate endothelial progenitors. The dark cells were scarcely myoid, populated the episcleral perivascular niches and the scleral stroma, and were equipped with lipid storage areas such as lamellar bodies and lipid droplets (LDs). Previously, unreported intranuclear LDs were found in these cells, which is characteristic of an HSC population. It appears that the human scleral stroma is a niche harbouring TC-like cells with immune and HSC phenotypes, and the mere presence or characteristics of telopodes are not enough to differentiate them.

Stromal cells/telocytes and endothelial progenitors in the perivascular niches of the trigeminal ganglion.

Stromal cells/telocytes (SCs/TCs) were recently described in the human adult trigeminal ganglion (TG). As some markers are equally expressed in SCs/TCs and endothelial cells, we hypothesized that a subset of the TG SCs/TCs is in fact represented by endothelial progenitor cells of a myelomonocytic origin. This study aimed to evaluate whether the interstitial cells of the human adult TG correlate with the myelomonocytic lineage. We used primary antibodies for c-erbB2/HER-2, CD31, nestin, CD10, CD117/c-kit, von Willebrand factor (vWF), CD34, Stro-1, CD146, α-smooth muscle actin (α-SMA), CD68, VEGFR-2 and cytokeratin 7 (CK7). The TG pial mesothelium and subpial vascular microstroma expressed c-erbB2/HER-2, CK7 and VEGFR-2. SCs/TCs neighbouring the neuronoglial units (NGUs) also expressed HER-2, which suggests a pial origin. These cells were also positive for CD10, CD31, CD34, CD68 and nestin. Endothelial cells expressed CD10, CD31, CD34, CD146, nestin and vWF. We also found vasculogenic networks with spindle-shaped and stellate endothelial progenitors expressing CD10, CD31, CD34, CD68, CD146 and VEGFR-2. Isolated mesenchymal stromal cells expressed Stro-1, CD146, CK7, c-kit and nestin. Pericytes expressed α-SMA and CD146. Using transmission electron microscopy (TEM), we found endothelial-specific Weibel-Palade bodies in spindle-shaped stromal progenitors. Our study supports the hypothesis that an intrinsic vasculogenic niche potentially involved in microvascular maintenance and repair might be present in the human adult trigeminal ganglion and that it might be supplied by either the pial mesothelium or the bone marrow niche.

The molecular phenotypes of ureteral telocytes are layer-specific.

Telocytes (TC) are the delicate interstitial (stromal) cells defined by their long, thin and moniliform processes termed telopodes. Numerous studies determined that different subsets of telocytes populate almost all tissues and attempted to relate these subsets to various functions, from cell signaling to tissue repair and regeneration. Extremely few studies addressed the urinary tract though few data on the molecular pattern of the urinary TCs actually exist. We therefore hypothesized that subsets of urinary TCs co-localize within the human ureter and we aimed at performing an immunohistochemical study to evaluate the tissue-specific molecular pattern of TCs. On sample tissues of proximal ureter drawn from ten human adult patients during surgery were applied primary antibodies against CD34, CD105, von Willebrand Factor, the heavy chain of smooth muscle myosin (SMM) and c-erbB-2. The molecular pattern indicated three different subsets of ureteral TCs which are neither endothelial nor epithelial in nature: (a) type I: the CD34-/CD105+ TCs of the superficial layer of lamina propria; (b) type II: the CD34+/CD105± myoid TCs of the deep layer of lamina propria and (c) type III: the CD34+/CD105+ perivascular TCs. Although apparently different, all these subsets of TCs could belong to the stem/progenitor niche of the ureter.

Subsets of telocytes: Myocardial telocytes.

Telocytes (TCs) are morphologically defined as small-sized cells with long, thin, moniliform processes called telopodes (Tps). Numerous papers imply that TCs are a distinctive cell type, and that transmission electron microscopy (TEM) is the gold standard tool for their identification. We aimed to reproduce previous studies on myocardial TCs to check their validity. For this purpose we performed an immunohistochemical study on human cardiac samples from six autopsied donor cadavers, using antibodies against CD10, CD31, CD34, CD146, Ki67, alpha-smooth muscle actin (α-SMA), Platelet-Derived Growth Factor Receptor-alpha (PDGFRα) and laminin. Additionally we performed a TEM study on cardiac samples from three human autopsied donor cadavers and five adult Sprague-Dawley rats. We found endothelial cells (ECs), cords, and filopodia-projecting endothelial tip cells (ETCs) that expressed CD10, CD31, CD34, CD146, and PDGFR-α. Often, endothelial cells closely neighbored the sarcolemmal basal laminae. Endothelial progenitor cells, as well as nascent capillaries, were CD31+/CD34+. Proliferative endothelial cells expressed Ki67. In larger vessels we found pericytes that expressed CD146 and α-SMA; scarce α-SMA-expressing spindle-shaped cells lining cardiomyocytes were suggestive of a pericytic role in angiogenic sprout guidance. The TEM study showed that endothelial tubes are almost exclusively found in the narrow myocardial interstitia. ECs that built them up appeared identical to the cells that previous TEM studies have suggested to be myocardial telocytes. A subset of stromal cells with TC-like phenotype and telopodes-like processes actually seem to configure blood vessels, and therefore belong to the endothelial lineage. This study shows that data presented in previous studies on myocardial telocytes is not enough to allow the reproducibility of the results. At least a subset of cells considered to be TCs might belong to the endothelial lineage.

Altered Mitochondrial Anatomy of Trigeminal Ganglia Neurons in Diabetes.

Neurons from sensory ganglia are exposed to oxidative attack in diabetes. Altered mitochondrial morphologies are due to impaired dynamics (fusion, fission) and to cristae remodeling. This study aimed to evaluate using transmission electron microscopy mitochondrial changes in diabetic trigeminal ganglia suggestive for ignition of apoptosis, in absence of "classical" morphological signs of apoptosis. We used samples of trigeminal ganglia (from six type 2 diabetes human donors and five streptozotocin (STZ)-induced diabetic rats). In human diabetic samples we found three main distributions of mitochondria: (a) small "dark" normal mitochondria, seemingly resulted from fission processes; (b) small "dark" damaged mitochondria, with side-vesiculations (single- and double-coated), large matrix vesicles and cytosolic leakage of reactive species, mixed with larger "light" mitochondria, swollen, and with crystolysis; (c) prevailing "light" mitochondria. In STZ-treated rats a type (c) distribution prevailed, except for nociceptive neurons where we found a different distribution: large and giant mitochondria, suggestive for impaired mitochondrial fission, mitochondrial fenestrations, matrix vesicles interconnected by lamellar cristae, and mitochondrial leakage into the cytosol. Thus, the ultrastructural pattern of mitochondria damage in diabetic samples of sensory neurons may provide clues on the initiation of intrinsic apoptosis, even if the classical morphological signs of apoptosis are not present. Further studies, combining use of biochemical and ultrastructural techniques, may allow a better quantification of the degree in which mitochondrial damage, with membrane alterations and cytosolic leaks, may be used as morphological signs suggesting the point-of-no return for apoptosis. Anat Rec, 299:1561-1570, 2016. © 2016 Wiley Periodicals, Inc.

Regenerative potential of human schneiderian membrane: progenitor cells and epithelial-mesenchymal transition.

An innate osteogenic potential of the Schneiderian membrane (SM) is progressively assessed in studies ranging from non-human species to human subjects. It has relevance for endosteal placement and osseointegration. Nestin-expressing osteogenic progenitor cells are allegedly involved in bone formation and remodelling. Nestin phenotype was not assessed previously in human SM. We therefore aimed to fill that particular gap in the literature. Bioptic samples of human adult SM were obtained during surgery from eight adult patients, operated for non-malignant pathologies. Immunohistochemistry on paraffin-embedded tissue samples used primary antibodies against nestin, CD45, CD146, cytokeratin 7 (CK7), and alpha-smooth muscle actin (α-SMA). Nestin expression was consistently found in endothelial cells, and was scarcely encountered in pericytes, putative stromal stem/progenitor cells, as well as in glandular epithelial cells. Moreover, woven bone formation in the periosteal layer of the SM can also be regarded as evidence of the osteogenic potential of this membrane. Nestin and CD45 expression in cells of the primary bone supports the osteogenic potential of SM nestin-expressing cells and a possible involvement of hematopoietic stem cells in maxillary sinus floor remodeling. CD146, a known inducer of epithelial-mesenchymal transition (EMT), was expressed in epithelia, as was CK7. Isolated stromal cells were found expressing CD146, CK7 and α-SMA, suggesting that regenerative processes happening in the SM may also involve processes of EMT which generate stem/progenitor cells. This study provides additional evidence for the regenerative potential of the Schneiderian membrane and identifies potential roles for cells of its stem niche in osteogenesis.

Brown adipocytes, cardiac protection and a common adipo- and myogenic stem precursor in aged human hearts.

New data on adult stem cells (ASCs) are continuously added by research for use in regenerative medicine. However organ-specific ASC markers are incompletely explored. It was demonstrated that in non-cardiac brown adipose tissue (BAT) CD133+ cells differentiate in cardiomyocytes, and such BAT-derived cells induce bone marrow-derived cells into cardiomyocytes, thus being a promising source for cardiac stem cell therapy. During embryogenesis the subepicardial fat derives from BAT. Although it was not specifically investigated in human adult or aged hearts, it is actually known that metabolically active BAT can be found in many adult humans, is related to antiobesity effects, and it may derive from stem/progenitor cells. Stro-1 can safely identify in situ cardiac stem cells (CSCs) with myogenic and adipogenic potential. It was therefore raised the hypothesis of subepicardial differentiation of CSCs in BAT in adult/aged hearts, which could be viewed, such as in infants, as a mechanism of protection. This could be determined by the reactivation of an embryologic differentiation pattern in which brown adipocytes and muscle cells derive from a common stem ancestor. Such quiescent common stem ancestors could be suggested in adult, or aged, human hearts, when subepicardial BAT is found, and if a Stro-1+/CD133+/Isl-1+ phenotype of CSCs is determined.

CD146- and CD105-positive phenotypes of retinal ganglion cells. Are these in situ proofs of neuronal regeneration?

The in vivo identity of stem cells is not yet clear. Numerous studies involve the perivascular niches as providers of stem cells during regenerative processes. CD146, in humans, as well as gicerin, at chicken, play roles in neuronal development and neurites extension. CD146 is a marker of stemness but also a pericytary marker. Stem cells in vascular niches can differentiate in neural cells. By applying CD146 and CD105 antibodies on human retinas from glaucomatous eyes, CD146-positive retinal ganglion cells (RGCs) were found, some being placed in perivascular positions; ongoing processes of neurites extension were related to these neurons. On other hand, RGCs were positively labeled by CD105 antibodies. These results support the hypothesis that in glaucoma eyes the CD146-positive RGCs result from regenerative processes driven by stem cells in the retinal perivascular niches. Further experiments are needed to evaluate whether CR146-positive neurons indicate also a physiological process of maintenance of retina.